AIM: To study the influences of P1 promoter activity of furin gene on the functions of hepatocytes in patients with liver cirrhosis.METHODS: The patients with liver cirrhosis of 180 cases were recruited.The single nucleotide polymorphism(SNP-229 C/T) in P1 promoter of furin gene was genotyped using competitively differentiated polymerase chain reaction.The relationships between the promoter activity based on genotyping and the serum levels of liver enzymes,total bilirubin,albumin and prothrombin were observed.RESULTS: The distribution frequencies of allele C and T were 75.3%(271/360) and 24.7%(89/360).Those of genotypes CC,CT and TT were 62.2%(112/180),26.1%(47/180) and 11.7%(21/180),respectively.The distribution frequencies of the genotypes were not related to the serum levels of major liver enzymes,albumin,total bilirubin and prothrombin,except for alkaline phosphatase and ?-glutamyl transferase.CONCLUSION: The activity of furin promoter exerts no effects on the main functions of hepatocytes,suggesting that furin may be a new therapeutic target for HBV infection.

A group of SARS-Iike coronaviruses(SL-CoV)have been identified in horseshoe bats.Despite SL-CoVs and SARS-CoV share identical genome structure and high-level sequence similarity,SL-CoV does not bind to the same cellular receptor as for SARS-CoV and the N-terminus of the S proteins only share 64% amino acid identity,suggesting there are fundamental differences between these two groups of coronaviruses.To gain insight into the basis of this difference,we established a recombinant adenovirus system expressing the S protein from SL-CoV(rAd-Rp3-S)to investigate its immune characterization.Our results showed that immunized mice generated strong humoral immune responses against the SL-CoV S protein.Moreover,a strong cellular immune response demonstrated by elevated IFN-γ and IL-6 levels was also observed in these mice.However,the induced antibody from these mice had weaker cross-reaction with the SARS-CoV S protein,and did not neutralize HIV pseudotyped with SARS-CoV S protein.These results demonstrated that the immunogenicity of the SL-CoV S protein is distinct from that of SARS-CoV,which may cause the immunological differences between human SARS-CoV and bat SL-CoV.Furthermore,the recombinant virus could serve as a potential vaccine candidate against bat SL-CoV infection.

Duck circovirus(DuCV),a potential immunosuppressive virus,was investigated in Southern China from March 2006 to December 2009 by using a polymerase chain reaction(PCR)based method. In this study,a total of 138 sick or dead duck samples from 18 different farms were examined with an average DuCV infection rate of～35%. It was found that ducks between the ages of 40～60 days were more susceptible to DuCV. There was no evidence showing that the DuCV virus was capable of vertical transmission. Farms with positive PCR results exhibited no regularly apparent clinical abnormalities such as feathering disorders,growth retardation or lower-than-average weight. The complete genomes of 9. strains from Fujian Province and 1 from Zhejiang Province were sequenced and analyzed. The 10 DuCV genomes,compared with others genomes downloaded from GenBank,ranged in size from 1988 to 1996 base pairs,with sequence identities ranging from 83.2% to 99.8%. Phylogenetic analysis based on genome sequences demonstrated that DuCVs can be divided into two distinct genetic genotypes,Group I(the Euro-USA lineage)and Group II(the Taiwan lineage),with approximately 10.0% genetic difference between the two types. Molecular epidemiological data suggest there is no obvious difference among DuCV strains isolated from different geographic locations or different species,including Duck,Muscovy duck,Mule duck,Cheery duck,Mulard duck and Pekin duck.

Objective To investigate if Rac1 GTPase activation plays an important role in hyperpermeability and tyresine phosphorylation of tight junction induced by vascular endothelial growth factor (VEGF) in glomertdar endothelial cells (GEnCs). Methods Primary cultured rat endothelial cells were used as experimental model. The effect of VEGF at different concentrations (5 or 50 μg/L) on endothelial permeability was investigated by transendothelial electrical resistance (TEER). The permeability of GEnCs transfected with wild type Rac1 (wtRacl) or dominant negative Racl (N17Rac1) was also detected. Immune precipitation and immune blotting were used to detect the tyrosine phosphor-occludin in GEnCs. Results VEGF at high concentration (50 μg/L) induced hyperpermeability in GEnCs (P<0.05). At the same time, GTP-binding and membrane-bound Racl GTPase significantly increased(P<0.01)in GEnCs. Tyrosine phosphor-occludin was also increased (P<0.05) under VEGF stimulation. However, transfection of GEnCs with N17Rac1 dramatically attenuated the effect of VEGF on tyrosine phospho-occludin and endothelial cell permeability. Conclusions Increased VEGF can induce hyperpermeability in glomerular endothelial cells, which is related to occludin tyrosine phosphorylation through Racl activation. It provides a framework for understanding the role of VEGF-induced Racl-phospho-occludin pathway in the integrity of barrier function in the diabetic milieu.

Objective To evaluate the effects of combination of dexmedetomidine and mild hypothermia on global cerebral ischemia-reperfusion (I/R) injury in neonatal rats.Methods Ninety-six neonatal Sprague-Dawley rats,aged 6-7 days,weighing 18-22 g,were randomly divided into 4 groups (n=24 each) using a random number table:I/R group,mild hypothermia group (group H),dexmedetomidine group (group D) and combination of dexmedetomidine and mild hypothermia group (group DH).Global cerebral ischemia was induced in rats anaesthetized with chloral hydrate by bilateral common carotid artery clamping (for 15 min) combined with hypotension followed by reperfusion.Dexmedetomidine 75 pg/kg was given intraperitoneally at 30 min before ischemia in D and DH groups,while the equal volume of normal saline was given in I/R and H groups.The temperature in the temporal muscle was maintained at 36.7-37.2℃ in I/R and D groups,and at 34.8-35.3℃ in H and DH groups.At 12,24 and 72 h of reperfusion,8 rats were randomly chosen in each group,and neurological deficit score (NDS) was determined.The animals were then sacrificed,and their brains were removed for determination of myeloperoxidase (MPO) activity (by spectrophotometry) and contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in brain tissues (using ELISA).Results Compared with I/R group,the NDS,MPO activity and contents of TNF-α and IL-6 were significantly decreased in the other three groups.The NDS,MPO activity and contents of TNF-α and IL-6 were significantly lower in DH group than in H or D group.Conclusion Dexmedetomidine can optimize cerebral protection providedby mild hypothermia against global cerebral I/R injury through inhibiting inflammatory responses in brain tissues of neonatal rats.

Objective To evaluate the precision of GFR using Gates method and compared with the results from renal pathological changes. Methods Twenty-seven patients whose 99Tcm-DTPA renograms had no obvious uptake phase were enrolled in Group A, and 27 patients whose 99Tcm-DTPA renograms had obvious uptake phase were enrolled in Group B. The measurement of GFR by Gates method was compared to the creatinine clearance measured and predicted by Cockroft-Gault (C-G), modification of diet in renal disease (MDRD) and SCr level. Renal pathological changes in two groups were compared using Pearson correlation and t test analysis. Results In Group A, GFR determined by Gates method did not show correlation with that estimated by C-G or 1/SCr (r = 0. 357,0. 376, both P ＞0.05), but was significantly correlated with GFR estimated by MDRD(r = 0. 440, P ＜ 0.05). In Group B, GFR determined by Gates method showed significantly correlation among GFR estimated by MDRD, C-G, and 1/SCr (r =0. 471, 0. 527,0. 452, all P ＜ 0.05). Renal tubulointerstitial damage score in Group A was higher than that in Group B (7.15±2.32, 3.70±3.06, t=4.66, P ＜0.001). Conclusions GFR determined by Gates method is less precise when 99Tcm-DTPA renogram has no obvious uptake phase than that when 99Tcm-DTPA renogram has obvious uptake phase. Renal tubulointerstitial damage is a strong indicator of no obvious uptake phase in 99Tcm-DTPA renogram.

Objective To evaluate the effect of wire-reinforced polyurethane epidural catheters on the success rate of epidural catheterization in the patients undergoing caesarean section.Methods A total of 182 pregnant patients,aged 25-43 yr,with body height of 145-178 cm,weighing 51-100 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective caesarean section under combined spinal-epidural anesthesia,were divided into 2 groups using a random number table:polyvinyl chloride epidural catheter group (group Ⅰ,n =94) and wire-reinfnrced polyurethane epidural catheter group (group 11,n=88).Spinal or epidural puncture was performed at L2,3 or L3,4 interspace,and the corresponding epidural catheter was inserted in each group aficr succcssful puncturc.Thc dcvclopment of difficult insertion,intravascular catheter insertion or paresthesia during puncture or insertion was defined as a failure of epidural catheterization.The occurrence of failed epidural catheterization was recorded.Results The failure rate of epidural catheterization was significantly lower in group 1Ⅱ than in gronp Ⅰ (P＜0.05).Conclusion Wire-reinforced polyurethane epidural catheters can raise the success rate of epidural catheterization in the patients undergoing caesarean section.

Itraconazole has been used to treat superficial candidal infections in China for 12 years with promising efficacy and safety. This article retrospectively reviewed literatures published in the mainstream journals in China with an attempt to find a reasonable therapy for Chinese populations.
Antifungal Agents ; therapeutic use ; Candidiasis ; drug therapy ; Dermatomycoses ; drug therapy ; Female ; Humans ; Itraconazole ; therapeutic use ; Male ; Retrospective Studies ; Stomatitis ; drug therapy ; microbiology ; Vaginitis ; drug therapy ; microbiology

Objective To study the clinical significance of the change of plasma BNP levels in patients with grades sepsis and its correlation with the left ventricular ejection fraction.Methods One hundred patients of sepsis who were admitted in intensive care unit of Taizhou hospital were selected.It was divided into four groups:30 patients with septic shock,40 patients with severe sepsis,30 patients with sepsis and 20 persons without disease as control group.The concentration of plasma BNP was determined using sandwich immunoflurescence,and ultrasonic cardiogram was used to evaluate heart function and clinical features in all groups.To compare with the differences of the concentration of plasma BNP and the correlation between the concentration of Plasma BNP and LVEF in all groups,the data of clinical features,28-day mortality,prognostic values of BNP and the length of stay (ROG) in ICU were collected and compared.Result Plasma BNP levels in patients with septic shock[ (976.3 ± 160.7) pg/ml] were obvious higher than severe sepsis[ (648.4 ± 267.3) pg/ml ],sepsis [ (217.2 ± 89.7) pg/ml ] and control group [ (50.3 ± 25.4)pg/ml] (P ＜0.01).LVEF in patients with septic shock [ (48.2 ±9.6)% ] was obvious lower than severe sepsis[ (52.8 ±9.4)% ],sepsis[ (61.3 ± 8.9)% ] and control group[ (66.4 ±9.3)% ] (P ＜0.05 or P ＜0.01).It appeared to be inverse relationship between LVEF and the plasma BNP levels (r =-0.876,-0.724,P ＜0.01).BNP levels were significantly higher in non-survivors compared with survivors[ (1367.6±506.4)pg/ml vs (420.3 ±82.6)pg/ml,P ＜0.01 ].The receiver operating characteristic (ROG) curves indicated that values of areas under the curve of plasma BNP levels for 28-days mortality were 0.918(P ＜0.01).Conclusion The concentration of plasma BNP in patients was different in different grades of sepsis.It appeared to be negative correlation between the concentration of plasma BNP and LVEF.Plasma BNP levels had predictive value to the patients with sepsis.

OBJECTIVE:To create an in vitro harvesting method of culturing a large number of adult bone marrow MSCs(BMSCs) DESIGN,TIME AND SETTlNG:The randomized, controlled study was performed at the Shanghai Institute of Digestive Surgery (Key Laboratory of Education Committee of Shanghai City),as well as Department of General Surgery and Organ Transplantation Center,Ruijin Hospital,Medical College.Shanghai Jiao Tong University from September 2005 to April 2006.MATERIALS:Bone marrow samples were collected from normal persons.who did bone marrow examination at the Department of Hematology,Ruijin Hospital,Medical College.Shanghai Jiao Tong University.Donors were volunteers who signed the informed consent.METHODS:Human BMSCs were harvested using Pemoll gradient centrifugation and adherence method.and then incubated in microcarrier cytodex3.Common monolayer polystyrene was incubated as controls.Cell phenotype and proliferative activity were tested utilizing flow cytometry and MTT.MAIN OUTCOME MEASURES:Collection.incubation,morphology of human BMSCs.and prolireration and cell cycle of human BMSCs on the cytodex 3 were measured.RESULlTS:Flow cytometry detection showed that the surface marker of human BMSCs on the cytodex3 was ldentical to that on the common monolayer polystyrene;BMSCs were positive for CD29,CD44 and CD105.but negative for CD14,CD34,CD45,VLA-1 and HLA-DR.MTT detection demonstrated that human BMSCs were in the adaptive phase at days 1-3.and entered logarithmic phase frOm day 3.No significant difference was detected in human BMSCs on the monolayer polystyrene and cytodex3(P＞0.05).On the monolayer polystyrene,human BMSCs entered degenerating stage from day 6,whereas on the cytodex3,human BMSCs were still in the logarithmic growth phase at day 9(P＜0.05).Flow cytometry detection confirmed that the cell cycle of human BMSCs was the same both on the monolayer polystyrene and cytodex3 (P＞0.05). CONCLUSION:Using cytodex3 culture technique,a large amount of human BMSCs can be obtained,and the proliferative activity of these BMSCs is good.